ES2832734T3 - Visual or related performance improvements - Google Patents

Abstract

A non-therapeutic method of improving the visual performance of a human subject without age-related macular degeneration in need of said improvement, the method comprising the step of administering to the subject an effective amount of a composition comprising mesozeaxanthin, lutein and zeaxanthin as a supplement food or food additive, and wherein performing the method improves contrast sensitivity under photopic conditions.

Description

DESCRIPTION

Visual or related performance improvements

Field of the invention

The present invention relates to a composition and a method for improving visual performance in a human subject and to a method of manufacturing the composition.

Background of the invention

The central retina, known as the macula, is responsible for color and the vision of fine details. A pigment, made up of the two dietary carotenoids, lutein (L) and zeaxanthin (Z) and a typically non-dietary carotenoid, mesozeaxanthin (MZ), accumulates in the marrow where it is known as macular pigment (MP). . MP is a blue light filter and a powerful antioxidant. And it is therefore believed to protect against age-related macular degeneration (AMD), which is now the most common recorded cause of blindness in the Western world. Several scientists have proposed that macular pigments can enhance visual performance (VP), but there does not appear to be any persuasive experimental evidence to support this hypothesis.

Compositions containing MZ have been disclosed as useful in the treatment of age-related macular degeneration (AMD) see, for example, US 6329432. Supplements each containing L, Z and MZ are known and sold for the intended purpose of treating and / or preventing eye disorders such as AMD. An example of such supplements is sold under the trademark MacuShield®, and contains the three carotenoids of MP L, Z, MZ in the amounts of 10 mg, 2 mg and 10 mg, respectively, per dose.

WO 03/063848 discloses the use of a compound, such as lutein, zeaxanthin, mesozeaxanthin or mixtures thereof for the manufacture of a composition for improving the visual performance of a subject in dark conditions. The document, however, is quite unusual in that it does not contain any experimental evidence or data to support the intended use. Therefore, experts in the field would be quite skeptical of the disclosure and certainly could not harbor any hope of success with it.

EP 1920711 discloses a method of evaluating visual performance which, in fact, involves measuring or determining the amount of macular pigment (such as lutein, zeaxanthin or mesozeaxanthin) present in the subject's eye (i.e. measuring the optical density of the macular pigment, MPOD). If the level of MPOD is low, the document suggests administering a composition comprising lutein or zeaxanthin, which is supposed to lead to an improvement in visual performance. However, the document does not disclose any actual experimental data showing that improving the level of macular pigment can lead to an improvement in visual performance. Therefore, again, experts in the field would treat the disclosure of the document with some caution and would not have any hope of success with it.

Studies have been published investigating whether macular pigment augmentation with lutein and zeaxanthin supplements improves visual performance: "The impact of macular pigment augmentation on visual performance in normal subjects: COMPASS", Nolan M. John et al., Vision Research 51 (2011), 459-469 and “Supplementation with the carotenoids lutein or zeaxanthin improves human visual performance”, Kvansakul, Jessica et al., Ophthalmic Physiology Opt. 2006, 26.

362-371.

Summary of the invention

The inventors have found that the consumption of a dietary supplement containing lutein alone has little effect on the PM of subjects showing an abnormally low concentration of PM in the central part of the retina. In contrast, consumption of a food supplement comprising MZ alone can return MP levels in the central part of the retina to substantially normal, but has little effect on MP levels outside the central part. Consumption of a combined supplement, which contains relatively high amounts of MZ, but also Z and L, can not only normalize MP levels in the central region of the retina, but also increase MP levels outside the central region of the retina. the retina.

For present purposes, the "central region" of the retina means the central part of the retina that has an eccentricity of 0.25 ° or less, as determined by optical coherence tomography (OCT) and / or fundus photography.

For the purposes of the present specification, a subject is considered to be AMD free if they have a value of 1-3 in the AREDs 11-stage maculopathy classification system (Age-Related Eye Disease Study) (Klein et al., 1991 Ophthalmology 98, 1128-1134).

The invention provides a non-therapeutic method of improving the visual performance of a human subject without age-related macular degeneration in need of such improvement, the method comprising the step of administering to the subject an effective amount of a composition comprising mesozeaxanthin, lutein and zeaxanthin. . The composition is administered orally, as a food supplement or food additive. The realization of method improves contrast sensitivity under photopic conditions.

An effective amount of the composition for a particular subject can easily be determined by routine, non-innovative trial and error, in view of the guidelines provided herein. Low doses can be administered initially and the dosage increased until an improvement in visual performance is detected. The subject's visual performance can be tested in any of a number of convenient methods, explained in detail below.

For present purposes, MZ is understood to refer to the compound (trans 3R, 3'S meso) -zeaxanthine, having the structure shown in Figure 1. Also included within the term "MZ" are esters of MZ, for example, acetate, laurate, myristate, palmitate, linoleate, linolenate and arachidonate esters, and esters with omega 3 fatty acids.

A human subject is considered not to be experiencing AMD if, after examination by a retinologist, there is no sign of any of the following features normally associated with AMD, including: soft drusen, hyper- and / or hypopigmentary changes in the macula (AMD premature) or geographic atrophy or choroidal neovascularization (advanced AMD).

The composition will preferably comprise MZ at a concentration of at least 0.001% w / w to 20% w / w. In one embodiment, a preferred MZ concentration may be in the range 3-10% w / w. However, those skilled in the art will appreciate that the precise concentration of MZ in the composition of the invention is not critical: a beneficial effect on the visual performance of the subject can be obtained by consuming higher doses of a composition comprising lower concentrations of MZ and vice versa. . A typical effective average daily dose of MZ to be consumed by a normal human adult subject will typically be in the range of 0.1 mg to 100 mg per day, more conveniently in the range of 1 to 50 mg per day, and preferably in the range 5-25 mg per day.

The composition may conveniently be in unit dose form, for example, as a tablet, capsule or the like. Conveniently, but not necessarily, the composition may be packaged in an aluminum blister, of the type known to those skilled in the art. Desirably, one or two of the doses are taken each day, with the amount of MZ in the doses being adjusted accordingly.

The composition comprises MZ, lutein, and zeaxanthin, which can be collectively referred to as macular carotenoids. Conveniently, though not necessarily, MZ will be present in the composition at a concentration greater than or the same concentration as lutein and zeaxanthin. The percentage of MZ or lutein in the composition can vary from 10% to 90% (of macular carotenoid pigment present in the formulation). The percentage of zeaxanthin can typically range from 5 to 45% (of macular carotenoid pigment in the formulation). A particularly favored composition has a MZ: lutein: zeaxanthin ratio of 10: 10: 2 (or 45%, 45%, 10%).

The three macular carotenoids can preferably be combined or manufactured as is in individual formulation. The composition of the invention can be in any formulation suitable for oral consumption by a human subject, including a tablet, capsule, gel, liquid, powder or the like. Macular carotenoids can be granulated, for example, as microcapsules prior to inclusion in the formulation. The composition may suitably comprise conventional diluents, especially vegetable oils such as sunflower, safflower, corn and rapeseed oils, excipients, thickeners and the like which are well known to those skilled in the art. Said substances include calcium and / or magnesium stearate, starch or modified starch.

Other conventional formulating agents may be present in the composition, including any one or more of the following non-exclusive list: acidity regulators; anti-caking agents (e.g. sodium aluminosilicate, calcium or magnesium carbonate, calcium silicate, sodium or potassium ferrocyanide), antioxidants (e.g. vitamin E, vitamin C, polyphenols), colorants (e.g. artificial colorants such as FD & C Blue # 1, Blue # 2, Green # 3, Red # 40, Red # 3, Yellow # 5 and Yellow # 6; and natural colorants such as caramel, achiote, cochineal, betanin, turmeric, saffron, paprika, etc.); color retention agents; emulsifiers; flavorings; aroma enhancers; preservatives; stabilizers; sweeteners and thickeners.

The above-mentioned compositions containing MZ, lutein and zeaxanthin can be added to a preparation containing essential vitamins and minerals; for example, a once daily tablet / capsule that contains all the RDAs of the vitamins and minerals required by man; or diet products that are fortified by vitamins and minerals; or together with omega 3 fatty acids.

MZ-containing macular carotenoids can be supplied to chickens and their eggs can provide an excellent source of MZ for human consumption.

Visual performance

Visual performance is a state, situation or parameter, not an abnormality or disease. Therefore, there is a range of values in normal subjects without the presence of any underlying retinal or macular disease. However, like all other human situations, VP improvements are considered beneficial and desirable.

There are many different measures of "visual performance" known to those of skill in the art.

For present purposes, improving "visual performance" means producing a detectable improvement in one or more of the following in the subject: contrast sensitivity; visual acuity, preferably better visual acuity with correction; glare disability; glare nuisance; diffusion of eye light; photo-stress recovery; and S-cone sensitivity. Preferably, the improvement in visual performance created by consumption of the composition of the invention comprises an improvement in one or more of: contrast sensitivity, improved visual activity with correction or glare disability.

Preferably, the consumption of the composition of the visual performance parameters, more preferably in two or more and much more preferably a detectable improvement in three or more of the aforementioned visual performance parameters.

The various visual performance parameters listed above are described in more detail below.

(i) Contrast sensitivity function

Contrast is the difference in visual properties that make an object (or its representation in an image) distinguishable from other objects and the background. Visual perception of the real world, contrast is determined by the difference in color and brightness of the object and other objects within the same field of view. Contrast sensitivity is a measure of the subject's sensitivity to changes in contrast; it is a measure of how much contrast is required to accurately detect a target as distinct from its background.

By altering the size (spatial frequency) of a target, and the brightness of the background, it is possible to test the contrast sensitivity function, which reflects much better the vision of the real world, where the most important determinants of vision are contrast, size and luminosity. Contrast sensitivity function can be assessed using the Functional Acuity Contrast Assay (FACT), which is designed to test contrast sensitivity at variable spatial frequency settings, as disclosed by Loughman et al., 2010 Vision Res. 50 , 1249-1256). Letter contrast sensitivity can be measured using the commercially available "Thomson Diagram".

(ii) Visual acuity

Visual acuity is a simple and intuitive way to assess visual performance. It is a useful measure of vision because it refers directly to the need for glasses (ie, whether an individual is long or short-sighted, the introduction of spectacle lenses typically creates a predictable improvement in visual acuity). Furthermore, it tends to be adversely affected by eye disease and therefore abnormal visual acuity can be a sign of abnormality development.

Despite widespread use and popularity, it is not the best technique for vision assessment because (a) it tends not to relate well to vision under conditions other than the brightly lit, high-contrast test environment, and (b) solely assesses performance at high spatial frequency (ie, small font size) at the end of the spectrum.

Best corrected visual acuity ("BCVA") is normally assessed using a high contrast letter diagram (close to 100%, ie, black letters on a white background), after the subject's vision has been corrected with corrective lenses. to the best possible level. The subject's task is to read the smallest possible font size that he can recognize. Visual performance is quantified using a conventional annotation (eg, Snellen annotation; where 20/20 or 6/6 vision is accepted as normal human vision). Improvements in BCVA imply a benefit in overall vision acuity.

(i¡¡) Glare disability

Glare disability is a term used to describe the degradation of visual performance normally caused by loss of image contrast on the retina. Glare disability is often caused, for example, by surface light reflections or bright light sources such as car headlights and is usually a consequence of increased direct light scattering within the eye. New bi-xenon high intensity discharge ("HID") car headlights contain more "blue" light and are often considered a cause of additional glare compared to older headlight sources.

This is of particular importance for macular pigment research because of the optical filtering properties of macular pigment. The macular pigment acts as a filter for short-wavelength light (blue). Its prereceptorial and central location facilitates optimization of visual performance with respect to glare because intraocular direct light scattering is dominated by short-wavelength light (blue).

Glare disability can be assessed using the functional acuity contrast test (FACT), as disclosed by Loughman et al., 2010 Vision Res. 50, 1249-1256.

(iv) Glare nuisance

Glare annoyance causes an instinctive desire to look away from a bright light source or difficulty observing a task. Refers to the sensation experienced when global illumination is too bright, for example in a snow field in bright sunshine.

Macular pigment has the ability to lessen the effects of glare nuisance because (a) it filters out the blue component that contains most of the energy; less light and less energy therefore reach the photoreceptors to affect performance and (b) macular pigment also has dichroic properties which means it has the ability to filter plane polarized light. Plane polarized light is light reflected from a surface (eg snow covered ground, water, etc.) into the eye. It is unidirectional so that energy is concentrated and therefore has an increased effect on vision. This is why skiers, fishermen and the like wear polarized sunglasses to reduce such annoying glare.

Glare nuisance is assessed using a nuisance rating scale as reported by Wenzel et al., 2006 Vision Res. 46, 4615-4622.

(v) Diffusion of eye light

The diffusion of ocular light is a parameter that is relatively new in clinical practice after being studied for many years in experimental settings. It refers to the part of the incident light that is scattered through the ocular environment and does not participate in the formation of the normal image on the retina. Instead, this light creates a more or less homogeneous haze over the image of the retina. Several pathologies are known to increase the diffusion of light from the retina considerably, which can lead to symptoms such as loss of contrast sensitivity, glare disability, and halos. This will reduce the quality of a patient's vision in their daily life, for example while driving at night and the recognition of a person in front of a light source, but it will have only a very limited effect on visual acuity measured during a ophthalmic exam.

As the macular pigment absorbs the dominant short wavelength scattered component, it has the ability to significantly reduce the amount of diffusion of the ocular light, and therefore further enriches the visual performance particularly in glare circumstances.

Ocular light diffusion is assessed using the Oculus C-Quant as reported by van Bree et al., 2011 Ophthalmology 118, 945-953.

(vi) Photo-stress recovery

The photo-stress recovery test is a method of evaluating visual performance by timing the recovery of visual function after adaptation to an intense light source. The test involves exposing the macula to a light source bright enough to whiten a significant portion of the visual pigments. Return to normal retinal function and sensitivity depends on regeneration of visual pigments, the test essentially provides an indirect assessment of macular function.

Photo-stress recovery is assessed using an automated macular photo-stress test using the Humphrey Perimeter as disclosed by Loughman et al., 2010 Vision Res. 50, 1249-1256.

(vii) S cone sensitivity

The S cones are the "blue" sensitivity cones, that is, their maximum sensitivity is at short wavelengths. Typically, a person with high levels of macular pigment would be expected to demonstrate low S-cone sensitivity, as the macular pigment is minimizing the amount of blue light that hits the photoreceptors. Combining an S-cone sensitivity test with a photo-stress test can provide information on the direct effects of macular pigment on the actual sensitivity of those cones most affected by glare.

S-cone sensitivity is evaluated using the short wavelength automated perimetry program (SWAP) on the Humphrey Perimeter as described by (Davison et al., Optom. Vis. Sci. 2011 vol. 88).

(viii) Evaluation of VP by a questionnaire

Another method of testing improvement in visual performance is the use of a questionnaire to assess the subject's own evaluation of their visual performance. In preferred embodiments of the invention, therefore, it is determined a detectable improvement in visual performance by an increased value on a subjective evaluation questionnaire after an appropriate period of weeks or months of composition consumption, compared to a control evaluation questionnaire completed before beginning composition consumption .

A suitable questionnaire is disclosed by Charalampidou et al., Arch. Ophthalmol. 2011 (May 9, Epub ahead of print), describing an unvalidated 30-part "Visual Function in Normal Subjects" questionnaire (VFNq30), which was designed to assess subjective improvement in visual performance. The design was based in part on a previously validated visual activities questionnaire (Sloane et al., "The Visual Activities Questionnaire: Developing an instrument for assessing problems in everyday visual tasks. Technical Digest, Noninvasive Assessment of the Visual System, Topical Meeting of the Optical Society of America 1992), but adapted to fit a normal, young and healthy population sample. This questionnaire allows the subject to quantify their visual performance using three different metrics: situational analysis (SA) which requires the subject to classify their visual performance in specified daily life situations; comparative analysis (CA) that requires the subject to compare their perceived visual performance with that of their peers / family / friends; the subject satisfaction value (SSS) that requires the subject to provide an overall estimate of your perceived quality of vision. Each of the three metrics above is calculated to give a value performance for five different functional aspects of your vision: acuity / spatial vision: glare disability; light / dark adaptation; daily visual tasks; and color discrimination.

Time to achieve a VP upgrade

Obviously, no measurable, discernible or detectable improvement in the visual performance of a subject is expected immediately after consuming the composition of the invention. The period of dietary supplementation required to produce a measurable improvement in visual performance will depend on several factors, including the average daily dose size of macular carotenoids in the subject prior to commencing the nutritional supplementation, the subject's general health, etc. Normally, it would be expected that nutritional supplementation with the composition of the invention would be required for at least 8 weeks, and more preferably at least 3 or 6 months before measuring one or more parameters of visual performance to test any improvement in it.

The subject may need to consume the active composition of the invention at least once a week, more usually at least 3 times a week and preferably daily.

Preferred embodiments

In one embodiment of the invention, the composition is consumed by subjects who are deficient in the amount of macular pigment in the central part of their macula. By way of explanation, the inventors have found that there is a proportion of the general population who cannot experience AMD (as defined herein), but who possess statistically significant lower levels of macular pigment in the center of the brain. macula determined by custom heterochromatic blink photometry (cHFP) using the Macular Densitometer ™. These subjects are described as having an atypical macular pigment distribution, referred to as "central depression". Using this technique, MP is psychophysically measured by HFP. HFP is based on the fact that MP absorbs blue light. The subject is asked to observe a target, within a test field, which alternates in a square wave push-pull between blue (460 nm) and green (550 nm) light, that is, flickering. The brightness of the blue light must be adjusted to achieve zero flicker, in other words, until the target is fixed. The ratio of the amount of blue light needed to achieve zero flicker in the fovea is compared to that needed in the parafovea (where the MP is assumed to be zero), the logarithm of which is known as the optical density. Using the densitometer, PM can be measured at five points across the macula; 0.25 °, 0.5 °, 1 °, 1.75 ° and 7 °. The HFP principle remains the same for each target. For those eccentricities of the retina outside the fovea, that is 0.5 °, 1 °, 1.75 ° and 7 °, the fixation point is located at the desired angular distance from a blinking disc. Three measurements are taken at each location and an average is calculated. To minimize error in HFP settings, care is taken to optimize the blink rate for each subject, otherwise known as the critical blink rate (CFF). The CFF is the frequency at which the subject can no longer perceive flicker on a 0.5 ° target at 550 nm. The CFF was determined using a cutoff method whereby the blink rate is progressively decreased (or increased), until the subject reports a change from blend to blink (or blink to blend). Subjects with an atypical macular pigment distribution ("central depression") have a MPOD at 0.5 ° eccentricity that is greater than or equal to the MPOD at 0.25 ° eccentricity.

In another embodiment the composition is consumed by subjects who have statistically normal levels of normal pigment.

Also disclosed is a method of manufacturing a composition for human consumption, the composition must be consumed by a human subject in order to improve visual performance, the method comprising the step of mixing an effective amount of MZ with a diluent, excipient or acceptable dietary vehicle. The method may further comprise adding lutein and / or zeaxanthin to the diluent, excipient, or carrier (or vice versa). Performance of the method will desirably result in the manufacture of a composition having the preferred characteristics set out above. The method may further comprise the step of packaging the composition in a container together with instructions for consuming the composition to achieve an improvement in visual performance. Conveniently, the composition can be packaged in unit dosage form, for example, as a plurality of tablets, capsules or pills that can be packaged singly (for example, in a tube) or they can be packaged individually (for example, in a blister pack).

A method of improving the visual performance of a human subject is also disclosed, the method comprising the steps of:

a) supplying a feed to laying birds, such as chickens or ducks, whose feed comprises MZ, to cause the birds to lay eggs comprising MZ;

b) collecting said eggs and supplying the eggs, or at least part of their yolk in edible form, to the subject.

Unprocessed whole eggs can be provided for the subject to cook. Alternatively, the eggs can be processed and at least part of the yolks thereof provided to the subject, the MZ content of the eggs being concentrated in the yolk. Processing may involve, for example, peeling, cooking and drying of the eggs.

Typically the composition will be consumed at least once a week, preferably at least twice a week, more preferably at least three times a week, and most preferably at least daily. In some embodiments, the composition can be consumed more than once per day (eg, once in the morning and once in the evening). Those skilled in the art will appreciate that the frequency of consumption can be adjusted to account for the carotenoid concentration of the macular pigment, especially mesozeaxanthin, present in the formulation. The method can be adjusted accordingly.

Performing the method of the invention, for a sufficient period of time (typically at least 8 weeks, preferably at least 3 months, more preferably more than at least 6 months and much more preferably for 12 months or more) will typically cause an increase in the macular pigment level in a subject.

The amount of increase in the level of macular pigment carotenoids in the subject that is achieved by consumption of the composition may depend, for example, on the level of macular pigment carotenoids present in the subject's eyes prior to commencing consumption of the composition. . As described above, the inventors have found that there is a proportion of the population (approximately 10% or so) in Ireland who have abnormally low levels of macular pigment and an abnormal distribution of carotenoid pigments within the macula and it is anticipated that similar subjects exist in other populations. Such persons can be expected to show a substantial increase in the level of macular pigment after long-term consumption (ie, 6 months or more) of the composition of the invention.

However, significantly and surprisingly, the inventors have also discovered that at least some visual performance parameters (eg, letter contrast sensitivity; glare disability) can be improved by consuming the inventive composition without a necessarily corresponding increase in the level of macular pigment.

In particular, the composition / method of the invention can produce a detectable improvement in the visual performance of a subject in different low light conditions. For example, the composition / method of the invention can produce an improvement in the visual performance of a subject in lighting conditions greater than 1 Cdm-2; more especially in photopic conditions (eg illumination levels greater than or equal to 3 Cdm-2).

More especially, the composition / method of the invention can produce an improvement in contrast sensitivity (CS). Suitable methods of measuring these visual performance parameters are known to those of skill in the art and are described in detail in this document. Typically, the composition / method of the invention will produce an improvement of at least 5%, preferably at least 8%, more preferably at least 10%, over the same parameter measured prior to consumption of the composition / method performance. the invention.

For the avoidance of doubt it is hereby explicitly stated that any feature of the invention described herein as preferable, advantageous, convenient, desirable, typical or the like may be present in any embodiments of the invention in isolation or in combination with any one. or more of such characteristics, unless the context dictates otherwise. Furthermore, the features described in relation to one aspect of the invention will apply equally to the other aspects of the invention, unless the context dictates otherwise.

The invention will now be further described by illustrative embodiment and with reference to the accompanying drawings, in which:

Figure 1 is a schematic representation of the structural formulas for lutein, zeaxanthin, and MZ; Figure 2 is a plot of visual acuity corrected for central macular pigment OD (arbitrary units) in a group of normal and mixed AMD subjects;

Figure 3 is a graph of macular pigment OD (at 0.25 ° eccentricity) versus time for subjects consuming various macular carotenoid compositions;

Figure 4 is a graph showing the measurement of macular pigment OD, at varying degrees of eccentricity, for particular subjects found to have atypical MPOD profiles, with a "central depression" (ie, lower levels of macular pigment in the center of the macula);

Figures 5 and 6 are plots of MPOD (at 0.25 ° and 0.50 ° eccentricity, respectively) versus time, for subjects receiving one of three different macular carotenoid formulations;

Figures 7-9 are plots of mean MPOD versus retinal eccentricity for Groups 1-3, respectively (see Example 2), before and after an 8-week period of supplemental food intake with one of three different formulations of macular carotenoids; Y

Figure 10 is a plot of MPOD versus retinal eccentricity (mean of eight subjects; see example 5) before (circle symbols) or after (square symbols) 3 months of supplementation with a daily dose of 10 mg of L, 10 mg of Mz and 10 mg of Z.

Example 1:

Comparison of macular responses after supplementation with three different formulations of macular carotenoids

Subjects and recruitment

This study was conducted at the Institute of Vision Research, Whitfield Clinic, Waterford, Republic of Ireland. Seventy-one volunteer subjects came forward to participate in this study, which was approved by the local research ethics committee. The subjects were between the ages of 32 and 84 and were in good general health. The volunteers were divided into two groups: a group with AMD and a normal group. It was confirmed that 34 subjects had premature stage AMD in at least one eye (AMD group; classified and identified by the presence of drusen and / or pigmentary changes in the macula) and 37 subjects had no ocular pathology (normal group). Importantly, for the AMD group, significant efforts were made to identify patients with premature AMD who were not currently taking carotenoid supplements.

Study design and formulation

L = lutein MZ = mesozeaxanthin Z = zeaxanthin

This study was a single anonymous randomized controlled clinical trial of oral supplementation with three different formulations of macular carotenoids, as follows:

group 1: high-L group

(n = 24 [normal group = 12 and AMD group = 12];

L = 20 mg / day, Z = 2 mg / day);

group 2: mixed carotenoid group

(n = 24 [normal group = 13 and AMD group = 11];

MZ = 10 mg / day, L = 10 mg / day, Z = 2 mg / day);

group 3: high-MZ group

(n = 23 [normal group = 12 and AMD group = 11];

MZ = 18 mg / day, L = 2 mg / day of L).

All subjects were instructed to take one capsule (oil-based) per day with a meal for 8 weeks. Compliance was assessed by counting the tablets at each study visit.

Macular Pigment Optical Density Measurement (MPOD)

The spatial profile of MP was measured using custom heterochromatic blink photometry (cHFP) using the Macular Densitometer ™, a cHFP instrument that is slightly modified from a device described by Wooten and Hammond (2005 Optometry & Vision Science 82, 378-386) and by Kirby et al., (2010 Invest. Ophthalmol. Vis. Sci. 51,6722-6728).

Subjects were evaluated at baseline, two weeks, four weeks, six weeks, and eight weeks (V1, V2, V3, V4, and V5, respectively). MPOD was measured at the following eccentricities: at 0.25 °, 0.5 °, 1 °, 1.75 °, 3 ° but only the results at 0.25 °, the central part of the retina corresponding to the macula , are presented on this occasion.

Statistic analysis

The statistical software packages PASW Statistics 17.0 (SPSS Inc., Chicago, Illinois, USA) and R were used for the analysis and Sigma Plot 8.0 (Systat Software Inc., Chicago, Illinois, USA) were used. ) for graphic presentations. All the quantitative variables investigated showed a typical normal distribution. The 5% level of significance was used.

Results

MPOD and visual acuity

There was a positive and statistically significant relationship between central MPOD (at 0.25 °) and visual acuity with correction at baseline (r = 0.303, p = 0.008), as shown in Figure 2, which is a graph of acuity. Visual corrected against MPOD (arbitrary units), showing the point data for individual subjects in the two groups before supplementation with one of the three carotenoid formulations. This finding suggests that central MP is significantly and positively related to visual performance.

Increase in MPOD over time

At the eccentricity of 0.25 ° the initial MPOD was different for each group as follows; group 1: 0.42 ± 0.20 group 2: 0.44 ± 0.18 group 3: 0.49 ± 0.21, with a mean of all groups of 0.45 ± 0.20. To simplify the comparison, all groups are worked out to start at the mean value. The study demonstrated an increase in MPOD over time, as illustrated in Figure 3, which is a plot of MPOD at 0.25 ° eccentricity (arbitrary units) versus time (time points 1 to 5, corresponding to 0, 2, 4, 6 and 8 weeks, respectively). As can be seen in Figure 3, the greatest increase in central MPOD was achieved with formulation group 2 (MZ = 10 mg / day, L = 10 mg / day, Z = 2 mg / day), which was different in a different way. statistically significant for groups 1 and 3. There was no statistical difference between groups 1 and 3.

Conclusions

Surprisingly, the greatest effect on macular pigment was observed with the mixed carotenoid group (group 2) containing MZ at 10 mg, L at 10 mg, Z at 2 mg, while the results with the other two groups were very similar. . There appears to be synergy between MZ and L. That the high-MZ group (group 3) was able to increase MP shows that MZ can raise MPOD substantially without any contribution from the other carotenoids, but was less effective than MZ in combination. L.

Macular carotenoid supplementation in subjects with "central depression" in their spatial profiles of macular pigment

The central retina, known as the macula, is responsible for color and the vision of fine details (Hirsh and Curcio 1989; Vision Res. 29, 1095-1101). A pigment, composed of two dietary carotenoids, lutein (L) and zeaxanthin (Z), and a non-dietary carotenoid MZ (MZ), (Johnson et al., 2005 Invest. Ophtalmol. Vis. Sci .

46, 692-702) accumulates in the macula, where it is known as macular pigment (MP). MP is a blue light filter (Snodderly et al., 1984 Invest. Opthalmol. Vis. Sci. 25, 660-673) and a potent antioxidant (Khachik et al. 1997 Invest. Ophthalm. & Vis. Sci. 38, 1802-1811) and is therefore believed to protect against age-related macular degeneration (AMD), which is now the most common recorded cause of blindness in the Western world (Klaver et al., 1998 Arch. Ophthalmol. 116, 653-658).

MZ and Z are the predominant carotenoids in the foveal region, while L predominates in the parafoveal region (Snodderley et al., 1991 Invest. Ophthalmol. Vis. Sci. 32, 268-279). MZ concentration peaks centrally, with an MZ: Z ratio of 0.82 in the central retina (in 3 mm from the fovea) and 0.25 in the peripheral retina (11-21 mm from the fovea ) (Bone et al., 1997 Experimental Eye Research 64, 211-218). The MZ of the retina is produced mainly by isomerization of L of the retina, therefore, it represents lower relative levels of L and higher relative levels of MZ, in the central macula, and vice versa in the peripheral macula, and would also explain the cause of that MZ represents approximately one third of the total MP.

The concentration of MP varies greatly between individuals (Hammond et al., 1997 Journal of the Optical Society of America A-Optics Image Science & Vision, 14, 1187-1196). Atypical MP spatial profiles (ie "central depressions") are present in some individual MP profiles. Most importantly, these "central depressions" were confirmed to be real and reproducible features of the MP spatial profile, when measured using custom heterochromatic blink photometry (cHFP, a validated technique for measuring MP). The The importance of such variations, if any, in the spatial profile of MP (for example, the presence of a "central depression") is not yet known, but may be related to the presumed protective function of this pigment. For example, reduced MPOD in the center of the macula (ie, the presence of a "central depression") may be associated with an increased risk of developing AMD.

It has been shown that 12% (58 subjects from a sample database of 484 subjects) of the normal Irish population had a reproducible 'central depression' in their MPOD spatial profile and that such depression in the MP spatial profile is more common in elderly subjects and cigarette smokers (two of the established risk factors for AMD).

Example 2 Supplementation of formulations containing macular carotenoids to subjects with a "central depression" in their PM profile

The study described in this example was conducted with volunteer subjects from the aforementioned database (n = 58) in the "central depression study", who were identified and confirmed with "central depressions" in their spatial profile of MP (es i.e. MPOD at 0.5 degrees eccentricity was> MPOD at 0.25 degrees eccentricity, see Figure 4) and they were invited to participate in an 8-week supplemental trial with one of three different carotenoid formulations maculars (see below).

Methods

Subjects and study design

Fifty-eight subjects with "central depressions" in their MP spatial profile (identified from a primary MP database; n = 484) were invited to take part in the study. Of the 40 subjects who agreed to return for the trial, 31 were confirmed to still have a "central depression" (ie, MPOD at 0.5 degrees of eccentricity was> MPOD at 0.25 degrees of eccentricity) and they were therefore included in the 8-week supplementation trial.

All subjects signed an informed consent document and the experimental measures were in accordance with the Declaration of Helsinki. The study was reviewed and approved by the Research Ethics Committee, Waterford Institute of Technology, Waterford, Ireland. The inclusion criteria for participation in this study were as follows: MPOD at 0.5 degrees of eccentricity> MPOD at 0.25 degrees of eccentricity (ie, evidence of a "central depression" in the spatial profile of MP) ; without the presence of ocular pathology; visual acuity 20/60 or better in the study eye; not currently taking food supplements with L and / or Z and / or MZ.

Subjects were randomly assigned to one of the two groups as follows;

group 1: high L content group (n = 11), L = 20 mg / day, Z = 2 mg / day);

group 2: mixed carotenoid group (n = 10), MZ = 10 mg / day, L = 10 mg / day, Z = 2 mg / day);

group 3: high MZ group (n = 10), MZ 18 mg / day, L 2 mg / day).

All subjects were instructed to take one capsule per day with a meal for 8 weeks. MPOD was measured, including its spatial profile, that is, at 0.25 °, 0.5 °, 1 °, 1.75 °, 3 ° at baseline, four weeks, and 8 weeks.

Measurement of the optical density of the macular pigment

The spatial profile of MP was measured using cHFP using the Macular Densitometer ™, as described in Example 1. To measure the spatial profile of MP, measurements were made at the following degrees of eccentricity of the retina: 0.25 °, 0 , 5th, 1st, 1.75th, 3rd and 7th (the reference point) obtained using the following dimensioned target diameters; 30 minutes 1st, 2nd, 3.5th, 1st and 2nd.

Statistic analysis

The statistical software package PASW Statistics 17.0 (SPSS Inc., Chicago, Illinois, USA) was used for the analysis and Sigma Plot 8.0 (Systat Software Inc., Chicago, Illinois, USA) was used for graphic representations. All the quantitative variables investigated showed a typical normal distribution. The means ± SD are presented in the text and tables. Statistical comparisons of the three different intervention groups were made, at baseline, using independent sample t-tests and chi-square analysis, as appropriate. The 5% level of significance was used throughout the analysis.

Results

Change in MPOD over the 8 week supplemental contribution period

MPOD repeated measures ANOVA was performed, for all measured retinal eccentricities (i.e., at 0.25 °, 0.5 °, 1.0 °, 1.75 °, and 3 °) at time (es that is, about the study period [start, 4 weeks and 8 weeks]), using a general linear model strategy, with a factor between subjects: treatment (group 1, group 2, group 3) and age as a covariate. Figures 5 and 6 show the change in MPOD during the course of the test for measurements at 0.25 ° and 0.5 ° eccentricity, respectively. Table 1 presents the repeated measures ANOVA results for each group separately and for each degree of eccentricity of the retina. As seen in this table, the increase in MPOD at 0.25 ° and 0.5 ° was statistically significant in group 2 (ie, the mixed carotenoid group). Also, a significant increase in MPOD at 0.25 ° was observed in group 3 (ie, high MZ group). It should be noted that only the 0.25 ° increase in MPOD in group 2 remains significant after Bonferroni correction for multiple trials.

The change in the spatial profile of MPOD for each of the groups 1-3 is illustrated in Figures 7-9, respectively.

Conclusions

Only the two formulations containing MZ were able to correct the "central depression" and increase the MP. Surprisingly, and contrary to expectations, the formulation containing L, but without MZ, had no effect on MPOD at any eccentricity.

The formulation containing mixed carotenoids (group 2) obtained a superior effect since it increased the MP significantly at both eccentricities of 0.25 and 0.5. This is consistent with the result of subjects who were supplemented with all three carotenoids without a central depression at baseline (see Example 1), that is, maximal response was observed using a supplement each containing MZ, L, and Z.

Table 1. Average MPOD values in each degree of eccentricity for all subjects according to the group and per visit___________________________________________________________________________________________ MPOD Group Start 4 weeks 8 weeks Time interaction (p-value) Group 1 0.25 0.45 ± 0, 20 0.48 ± 0.22 0.49 ± 0.21 0.112

Group 1 0.5 0.45 ± 0.23 0.46 ± 0.18 0.46 ± 0.23 0.509

Group 1 1 0.20 ± 0.18 0.27 ± 0.15 0.25 ± 0.13 0.234

Group 1 1.75 0.15 ± 0.09 0.15 ± 0.09 0.15 ± 0.09 0.986

Group 1 3 0.15 ± 0.11 0.16 ± 0.09 0.11 ± 0.08 0.265

Group 2 0.25 0.41 ± 0.27 0.50 ± 0.27 0.59 ± 0.30 0.000

Group 2 0.5 0.44 ± 0.26 0.46 ± 0.28 0.52 ± 0.28 0.016

Group 2 1 0.26 ± 0.23 0.29 ± 0.15 0.34 ± 0.10 0.417

Group 2 1.75 0.18 ± 0.10 0.19 ± 0.06 0.22 ± 0.06 0.218

Group 2 3 0.16 ± 0.12 0.14 ± 0.06 0.19 ± 0.11 0.448

Group 3 0.25 0.48 ± 0.16 0.55 ± 0.19 0.57 ± 0.18 0.005

Group 3 0.5 0.48 ± 0.15 0.48 ± 0.17 0.50 ± 0.15 0.786

Group 3 1 0.32 ± 0.12 0.31 ± 0.13 0.34 ± 0.12 0.596

Group 3 1.75 0.11 ± 0.09 0.12 ± 0.07 0.13 ± 0.08 0.743

Group 3 3 0.12 ± 0.08 0.15 ± 0.07 0.15 ± 0.07 0.522

The values represent the mean ± standard deviation; n = 31; MPOD = macular pigment optical density; 0.25 ° = MPOD of measurement at 0.25 ° of eccentricity of the retina; 0.5 ° = MPOD of measurement at 0.5 ° of eccentricity of the retina; 1.0 ° = MPOD of measurement at 1.0 ° of eccentricity of the retina; 1.75 ° = MPOD of measurement at 1.75 ° of eccentricity of the retina; 3 ° = MPOD of measurement at 3.0 ° of eccentricity of the retina; group 1: high-L group; group 2: group of combined carotenoids; group 3: high MZ group; p-values represent repeated measures ANOVA for all 3 study visits (within-subject effects), with Greenhouse-Gesser correction for absence of sphericity as appropriate.

Example 3 Comparison of visual performance in subjects with AMD in premature phase after supplementation with three different formulations of macular carotenoids.

Subjects and recruitment

This study was conducted with 72 subjects, many with premature AMD. For details see example 1.

Study design and formulation

The subjects were divided into 3 groups (20-27 subjects) and the following supplementary inputs were administered:

group 1: L = 20; Z = 2 mg / day

group 2: L = 10; MZ = 10; Z = 2 mg / day

group 3: MZ = 17-18; L = 2-3; Z = 2 mg / day

These formulations, dissolved in 0.3 ml of vegetable oil, were administered in soft gel capsules.

Visual performance was measured using techniques previously described at baseline and 3 and 6 months after supplementation. Statistical analyzes were performed using a t-test for paired data. Significant values were considered as P <0.05. Results are only given when at least one group was statistically significant.

Results:

As there are no statistically significant improvements detected in PV after 3 months of treatment, this time only the results for 6 or 12 months are presented (below):

1. Initial comparison between groups:

Table 2 Initial comparison

Variable Group 1: Group 2: Group 3: p 20 mg L; 2 mg Z 10 mg MZ; 10 mg L; 2 mg Z 18 mg MZ; 2 mg L

N 23 27 22

Age 67 ± 8 64 ± 9 72 ± 10 0.014 MPOD 0.25 ° 0.412 ± 0.19 0.482 ± 0.21 0.475 ± 0.20 0.411 BCVA 92 ± 21 97 ± 10 94 ± 8 0.362

The groups were statistically comparable at baseline with respect to MP and vision (assessed by best acuity with correction, "BCVA"). There was a significant difference between the groups at baseline for age between group 3 and the other two groups. Group 1 and group 2 were statistically similar with respect to age.

2. Better visual acuity with correction (BCVA)

There was an initial correlation (before supplementation) of a positive and statistically significant relationship between central MPOD (0.25) and BCVA, importantly this is in the AMD population (r - = 0.368, p = 0.002) . There were no statistically significant changes in BCVA in either group after 3 and 6 months.

A computer generated LofMAR test chart (Test Chart 2000 Pro; Thomson Software Solutions) was used to determine BCVA at a viewing distance of 4 m, using a set of Sloan ETDRS letters. BCVA was determined as the average of three measurements, with changes in letter and line facilitated by the pseudo-randomization characteristic of the computer program. Best corrected visual acuity was recorded using a letter rating visual acuity rating, with 20/20 (6/6) visual acuity assigned a value of 100. Best corrected visual acuity was scored relative to this value , with each correctly identified letter assigned to a face value of one, so that, for example, a BCVA of 20/20 1 (6/6 + 1) equals a value of 101 and 20 / 20-1 (6 / 6-1) to 99.

3. MPOD response

The following table 3 presents the PM data for each group and for each eccentricity measured, at the beginning, six and twelve months after the supplementation with macular carotenoids.

A statistically significant increase in MPOD was observed at 12 months only in groups 2 and 3, who received the MZ-containing supplement.

Table 3

Group MPOD MPOD MPOD p Initial 6 months 12 months

0.25 0.25

1: 0.42 ± 0.19 0.51 ± 0.20 0.57 ± 0.30 0.148 2: 0.48 ± 0.22 0.58 ± 0.21 0.63 ± 0.19 0.001 3: 0.52 ± 0.20 0.58 ± 0.22 0.57 ± 0.20 0.022 0.5 0.5 p 1: 0.32 ± 0.19 0.42 ± 0.18 0.46 ± 0 .27 0.126 2: 0.39 ± 0.19 0.50 ± 0.18 0.52 ± 0.19 0.001 3: 0.41 ± 0.19 0.46 ± 0.19 0.45 ± 0.20 0.034

1.0 1.0 p 1: 0.22 ± 0.11 0.31 ± 0.15 0.32 ± 0.17 0.213 2: 0.25 ± 0.12 0.36 ± 0.17 0.37 ± 0.18 0.001 3: 0.26 ± 0.15 0.32 ± 0.14 0.33 ± 0.16 0.025

1.75 1.75 p 1: 0.13 ± 0.10 0.18 ± 0.11 0.20 ± 0.10 0.114 Group MPOD MPOD MPOD p

Initial 6 months 12 months

2: 0.14 ± 0.10 0.22 ± 0.12 0.24 ± 0.11 <0.001

3: 0.13 ± 0.11 0.21 ± 0.12 0.19 ± 0.10 0.063

4. Letter contrast sensitivity (Thomson diagram)

Table 4A presents the data for letter contrast sensitivity at baseline and six months after supplementation with macular carotenoids. Measurements were made at 1.2, 2.4, 6.0 and 9.6 cpd. There was a statistically significant improvement only in group 2 (10 mg MZ; 10 mg L; 2 mg Z) at 1.2, 2.4 and 9.6 cpd and not at all in the other two groups. This shows a vastly greater effect in group 2.

Table 4A:

Group Contrast sensitivity of letters Contrast sensitivity of letters p

Initial Six months

1: 1.2 cpd 1.2 cpd

1.68 ± 0.34 1.75 ± 0.30 0.091

2: 1.63 ± 0.24 1.80 ± 0.25 0.013

3: 1.68 ± 0.37 1.63 ± 0.25 0.322

2.4 cpd 2.4 cpd p

1: 1.60 ± 0.33 1.66 ± 0.34 0.17

2: 1.59 ± 0.29 1.72 ± 0.33 0.049

3: 1.61 ± 0.35 1.63 ± 0.32 0.6

9.6 cpd 9.6 cpd p

1: 1.1 ± 0.36 1.04 ± 0.41 0.194

2: 0.97 ± 0.32 1.11 ± 0.46 0.043

3: 0.94 ± 0.37 0.95 ± 0.43 0.901

Table 4B shows the letter contrast sensitivity (CS) at baseline and 12 months, for each of five spatial frequencies (1.2 - 15.15 cpd).

Table 4B. Mean values ± sd of contrast sensitivity of letters CS at baseline at 12 months.

Abbreviations: cpd = cycles per degree

At 12 months, the results were similar at 6 months because letter contrast sensitivity increased in all groups for large objects (1.2 and 2.4 cpd), but only in groups 2 and with smaller objects. (6.0-15.5 cpd).

Table 4C presents the relationship between the observed changes in MPOD (at 0.25 ° eccentricity) and the observed changes in letter CS at 1.2 cpd. It should be noted that there were no statistically significant relationships between the change in MP and the change in letter CS, at any spatial frequency.

Table 4C.

Photographs of the colored background

Background colored photographs were taken at each study visit using a Zeiss VisuCam ™ (Carl Zeiss Meditec AG, Jena, Germany) and stereoscopically graded at the Ocular Epidemiology Reading Center at the University of Wisconsin, USA. graduated using a modified version of the Wisconsin Age-Related Maculopathy Grading System. Premature AMD was defined as the presence of drusen and / or pigmentary changes in at least one eye, confirmed by a local ophthalmologist in collaboration with University of Wisconsin graders. Each fundus photograph was evaluated, lesion by lesion, to determine the maximum drusen size, type, area and pigmentary abnormalities of the retina. The overall findings were presented on an 11-stage AMD severity scale. A change of two or more stages along the severity scale was defined as clinically significant. Graduated photographs were obtained for the baseline and 12-month visits.

At baseline, there was no significant difference between the groups regarding the grade of AMD (p = 0.679) [Table 4D]

Table 4D. Grading of AMD for whole groups and subgroups at baseline.

Grade Complete group (n = 72) Group 1 (n = 23) Group 2 (n = 27) Group 3 (n = 22) Sig. 1-3 16 22.2% 7 30.4% 6 22.2% 3 13.6% 0.679

Changes in grade of AMD between baseline and 12 months for each of the three groups are summarized in Table 4E. A change in the negative direction (i.e., -1, -2) indicates progression along the AMD severity scale, while positive integers indicate regression (improvement) along the AMD severity scale. Between baseline and 12 months, there was no statistically significant difference between treatment groups with respect to change in severity of AMD (p = 0.223, Pearson's chi-square test).

Table 4E. Change in grade of AMD (11-stage scale) between baseline and 12 months

Group n -2 -1 0 1 2 Sig.

1 16 1 6% 1 6% 10 63% 3 19% 1 6% 0.223

Total 54 (100%) 4 (7%) 9 (17%) 28 (52%) 9 (17%) 4 (7%)

Abbreviations: n = number of subjects; a negative value indicates disease progression; a positive value indicates regression of the disease; 0 = no change in degree

It should be noted that Table 4E shows that 86% of the subjects did not show clinically significant changes in their AMD status between baseline and 12 months, showing a 7% deterioration and 7% showing an improvement (note: a change in the grade of two or more has been accepted as clinically significant).

Discussion

The most interesting results were for letter contrast sensitivity. This test is performed only in daylight and test letters of different sizes. The results at 6 months and 12 months were similar. There was no correlation between the increase in MP and the increase in this parameter indicating neurophysiological effects of macular carotenoids.

There was no significant change in the grade of AMD from baseline. Therefore, changes in contrast sensitivity were not related to the effects on AMD pathology.

5. Contrast sensitivity at night (assessed on the FACT device)

The following table 5 presents logarithmic contrast sensitivity data evaluated for the night, at the beginning and six months after the supplementation with macular carotenoids. Measurements were made at 1.5, 3.0, 6.0, 12 and 18 cpm. The statistically significant improvement in this VP measure was present only in group 2 at 1.5, 3.0 cpd and in group 1 at 1.5 cpd, which shows a greater effect than group 2.

Table 5:

Group Contrast sensitivity at night Contrast sensitivity at night

Initial Six months ^P 1.5 cpd 1.5 cpd

1: 1.53 ± 0.29 1.67 ± 0.26 0.124 2: 1.51 ± 0.27 1.66 ± 0.3 0.028 3: 1.44 ± 0.29 1.45 ± 0.34 0.911 Group 3.0 cpd 3.0 cpd p 1: 1.52 ± 0.25 1.8 ± 0.28 0.001 2: 1.62 ± 0.34 1.75 ± 0.41 0.01 3: 1 .55 ± 0.40 1.6 ± 0.41 0.585

6. Contrast sensitivity during the day (evaluated on the FACT device)

The following table 6 presents the logarithmic contrast sensitivity data evaluated for the day at the beginning and six months after the supplementation with macular carotenoids. Measurements were made at 1.5, 3.0, 6.0, 12 and 18 cpm. Statistically significant improvement in this VP measure was present in group 2 at 1.5, 3.0 and 18 cpd and in group 1 to 1.5 cpd, which shows a greater effect in group 2.

Table 6:

Contrast sensitivity by day Contrast sensitivity by day _Group p Initial Six months

1.5 cpd 1.5 cpd

1: 1.41 ± 0.16 1.57 ± 0.26 0.03 2: 1.48 ± 0.23 1.6 ± 0.28 0.034 3: 1.41 ± 0.13 1.5 ± 0 .28 0.238

3.0 cpd 3.0 cpd p 1: 1.67 ± 0.21 1.75 ± 0.21 0.17 2: 1.7 ± 0.33 1.81 ± 0.34 0.018 3: 1.72 ± 0.18 1.77 ± 0.29 0.46

18 cpd 18 cpd p 1: 0.62 ± 0.4 0.56 ± 0.41 0.497 2: 0.65 ± 0.38 0.77 ± 0.5 0.015 3: 0.57 ± 0.4 0, 62 ± 0.43 0.704

7. Contrast sensitivity at night in the presence of glare (evaluated on the FACT device)

The following table 7 presents the logarithmic data of contrast sensitivity at night in the presence of glare at baseline and six months after supplementation with macular carotenoids. Measurements were made at 1.5, 3.0, 6.0, 12 and 18 cpd. There was a statistically significant improvement in this PV only in group 2 at 18 cpd.

Table 7

Group Contrast sensitivity at night Contrast sensitivity at night p with glare with glare

Initial Six months

18 cpd 18 cpd

1: 0.34 ± 0.16 0.34 ± 0.16 0.136 2: 0.36 ± 0.13 0.47 ± 0.34 0.038 3: 0.36 ± 0.22 0.32 ± 0.08 0.588

8. Contrast sensitivity during the day in the presence of glare (evaluated on the FACT device)

The following table 8 presents the logarithmic contrast sensitivity data for the day in the presence of glare, at the beginning and six months after the supplementation with macular carotenoids. Measurements were made at 1.5, 3.0, 6.0, 12 and 18 cpd. The statistically significant improvement in this VP measure was present in group 2 at 1.5, 3.0, 6.0 and 18 cpd and in group 1 at 1.5, 3.0 and 6.0 cpd and in group 3 to 6 cpd, showing a higher effect in group 2.

Table 8

Contrast sensitivity by day Contrast sensitivity by day

with glare with glare P Initial Group Six months

1.5 cpd 1.5 cpd

1: 1.5 ± 0.25 1.63 ± 0.21 0.001 2: 1.43 ± 0.25 1.68 ± 0.24 0.002 3: 1.42 ± 0.38 1.46 ± 0.36 0.351

3.0 cpd 3.0 cpd p 1: 1.68 ± 0.22 1.85 ± 0.22 0.006 2: 1.71 ± 0.25 1.84 ± 0.25 0.007 3: 1.67 ± 0 .35 1.71 ± 0.43 0.542

6.0 cpd 6.0 cpd p 1: 1.46 ± 0.42 1.85 ± 0.22 <0.001 2: 1.46 ± 0.47 1.84 ± 0.25 <0.001 3: 1.36 ± 0.42 1.71 ± 0.43 0.001

18 cpd 18 cpd p 1: 0.64 ± 0.46 0.59 ± 0.43 0.642 2: 0.53 ± 0.32 0.67 ± 0.51 0.018 3: 0.7 ± 0.47 0, 66 ± 0.45 0.609

9. Contrast sensitivity and glare disability between initial measurement and 12 months Data on contrast sensitivity (CS) and glare disability (GD) in mesopic (at night) and photopic (daytime) conditions, at baseline and 12 months, are presented in Tables 9-12.

Table 9. Logarithm of CS at baseline and 12 months later under mesopic conditions (FACT device)

Group CS 1.5 cpd v1 CS 1.5 cpd v4 p

Group 1 1.59 ± 0.28 1.80 ± 0.22 0.007 Group 2 1.60 ± 0.27 1.76 ± 0.24 0.047 Group 3 1.53 ± 0.39 1.73 ± 0.25 0.124

CS 3 cpd v1 CS 3 cpd v4

Group 1 1.61 ± 0.25 1.82 ± 0.22 0.007 Group 2 1.68 ± 0.34 1.80 ± 0.26 0.058 Group 3 1.62 ± 0.42 1.85 ± 0.40 0.175

CS 6 cpd v1 CS 6 cpd v4

Group 1 1.18 ± 0.38 1.24 ± 0.53 0.521 Group 2 1.27 ± 0.40 1.38 ± 0.44 0.278 Group 3 1.20 ± 0.44 1.46 ± 0.50 0.060

CS 12 cpd v1 CS 12 cpd v4

Group 1 0.65 ± 0.14 0.79 ± 0.43 0.224 Group 2 0.67 ± 0.26 0.79 ± 0.24 0.080 Group 3 0.76 ± 0.25 0.89 ± 0.36 0.177

CS 18 cpd v1 CS 18 cpd v4

Group 1 0.40 ± 0.25 0.32 ± 0.08 0.207 Group 2 0.32 ± 0.07 0.36 ± 0.26 0.332 Group 3 0.36 ± 0.15 0.39 ± 0.24 0.476 Abbreviations: FACT = functional acuity contrast test; CS = contrast sensitivity; cpd = cycles per degree; v1 = initial visit; v4 = 12 month visit

Table 10. Logarithm of CS at baseline and 12 months under photopic conditions (FACT device)

Group CS 1.5 cpd v1 CS 1.5 cpd v4 p

Group 1 1.47 ± 0.25 1.63 ± 0.22 0.007 Group 2 1.56 ± 0.21 1.61 ± 0.24 0.478 Group 3 1.44 ± 0.22 1.63 ± 0.25 0.023

CS 3 cpd v1 CS 3 cpd v4

Group 1 1.70 ± 0.22 1.86 ± 0.11 0.002 Group 2 1.74 ± 0.33 1.86 ± 0.21 0.108 Group 3 1.78 ± 0.20 1.84 ± 0.24 0.402

CS 6 cpd v1 CS 6 cpd v4

Group 1 1.52 ± 0.30 1.59 ± 0.29 0.310 Group 2 1.52 ± 0.39 1.66 ± 0.39 0.064 Group 3 1.44 ± 0.45 1.62 ± 0.38 0.192

CS 12 cpd v1 CS 12 cpd v4

Group 1 1.01 ± 0.33 0.98 ± 0.35 0.709 Group 2 1.02 ± 0.36 1.21 ± 0.48 0.118 Group 3 0.99 ± 0.43 1.19 ± 0.48 0.164

CS 18 cpd v1 CS 18 cpd v4

Group 1 0.63 ± 0.39 0.54 ± 0.40 0.437 Group 2 0.59 ± 0.38 0.64 ± 0.48 0.687 Group 3 0.68 ± 0.48 0.76 ± 0.50 0.458 Abbreviations: FACT = functional acuity contrast test; GD = glare disability; cpd = cycles per degree; v1 = initial visit; v4 = 12 month visit

Table 11. Logarithm of GD at baseline and 12 months under mesopic conditions (FACT device)

Group GD 1.5 cpd v1 GD 1.5 cpd v4 p

Group 1 1.49 ± 0.37 1.52 ± 0.34 0.635 Group 2 1.44 ± 0.39 1.53 ± 0.35 0.365 Group 3 1.26 ± 0.44 1.53 ± 0.47 0.029

GD 3 cpd v1 GD 3 cpd v4

Group 1 1.57 ± 0.43 1.60 ± 0.32 0.728 Group 2 1.51 ± 0.38 1.70 ± 0.35 0.010 Group 3 1.39 ± 0.50 1.55 ± 0.49 0.346

GD 6 cpd v1 GD 6 cpd v4

Group 1 1.09 ± 0.37 1.04 ± 0.34 0.564 Group 2 1.18 ± 0.35 1.24 ± 0.43 0.581 Group 3 1.10 ± 0.40 1.20 ± 0.47 0.348 _____________________ GD 12 cpd v1___________________GD 12 cpd v4__________________________________ Group 1 0.66 ± 0.17 0.71 ± 0.18 0.343 Group 2 0.66 ± 0.17 0.80 ± 0.43 0.100 Group 3______________ 0.77 ± 0, 24 ______________________ 0.69 ± 0.22 _____________________ 0.115 __________ _____________________ GD 18 cpd v1___________________GD 18 cpd v4__________________________________ Group 1 0.34 ± 0.16 0.30 ± 0.00 0.336 Group 2 0.34 ± 0.10 0.39 ± 0.37 0.483 Group 3______________ 0.32 ± 0.08 ______________________ 0.36 ± 0.21 _____________________ 0.336 __________ Abbreviations: FACT = functional acuity contrast test; GD = glare disability; cpd = cycles per degree; v1 = initial visit; v4 = 12 month visit

Table 12. Logarithm of GD at baseline and 12 months under photopic conditions (FACT device) Group________________GD 1.5 cpd v1___________________ GD 1.5 cpd v4______________ p

Group 1 1.60 ± 0.25 1.76 ± 0.23 0.006 Group 2 1.53 ± 0.19 1.74 ± 0.22 0.002 Group 3 _______________ 1.51 ± 0.25 _____________________ 1.69 ± 0.42 ________________ 0.058 _____________________ GD 3 cpd v1 GD 3 cpd v4

Group 1 1.70 ± 0.26 1.89 ± 0.25 0.002 Group 2 1.78 ± 0.21 1.97 ± 0.18 0.001 Group 3_____________ 1.73 ± 0.20 1.84 ± 0.38 0.330 _____________________ GD 6 cpd v1 GD 6 cpd v4

Group 1 1.54 ± 0.38 1.64 ± 0.35 0.358 Group 2 1.56 ± 0.43 1.69 ± 0.34 0.087 Group 3_____________ 1.46 ± 0.47 1.71 ± 0.38 0.048 _____________________ GD 12 cpd v1 GD 12 cpd v4

Group 1 1.02 ± 0.42 1.05 ± 0.38 0.659 Group 2 0.97 ± 0.36 1.14 ± 0.35 0.169 Group 3 ______________ 1.00 ± 0.44 ____________________ 1.11 ± 0.43 _____________________ 0.320 __________ _____________________ GD 18 cpd v1___________________ GD 18 cpd v4__________________________________ Group 1 0.64 ± 0.45 0.67 ± 0.48 0.752 Group 2 0.54 ± 0.34 0.81 ± 0.51 0.071 Group 3 ______________ 0.75 ± 0 , 48 ____________________ 0.75 ± 0.52 _____________________ 0.993 __________ Abbreviations: FACT = functional acuity contrast test; GD = glare disability; cpd = cycles per degree; v1 = initial visit; v4 = 12 month visit

Discussion

The results at 12 months were similar to those at 6 months, because the results were variable and difficult to interpret. Under mesopic conditions (at night), contrast sensitivity increased only with large objects (1.5 and 3.0 cpd) in groups 1 and 2. For glare disability, group 1 did not change, while group 2 and 3 showed some change with large objects.

Under photopic conditions (daytime), groups 1 and 3 increased their contrast sensitivity only with large objects. With glare disability all groups increased only with large objects.

10. Changes in visual performance parameters and changes in MPOD

Table 13 presents the relationship between the changes observed in MPOD (at 0.25 ° eccentricity) and the changes observed in the parameters of visual performance, specifically CDVA and measurements of mesopic and photopic contrast sensitivity, and disability due to mesopic glare. and photopic, at 1.5 cpd. Notably, there were no statistically significant relationships between the change in MP and the change in visual performance in any of the groups (with the exception of a negative relationship between increases in MPOD and photopic CS at 1.5 cpd in the group 1 only.

Table 13.

Change in MPOD versus change in CDVA rp Group 1 -0.320 0.211 Group 2 -0.148 0.558 Group 3 -0.126 0.681 Change in MPOD versus change in mesopic CS 1.5 cpd rp Group 1 0.055 0.859 Group 2 -0.140 0.664 Group 3 0.041 0.906 Change in MPOD versus change in photopic CS 1.5 cpd r ____p__________ Group 1 -0.705 0.007 Group 2 -0.106 0.743 Group 3 -0.122 0.720 Change in MPOD versus change in mesopic GD 1.5 cpd rp Group 1 0.318 0.289 Group 2 -0.106 0.743 Group 3 0.388 0.238 Change in MPOD versus change in photopic GD 1.5 cpd rp Group 1 -0.262 0.388 Group 2 -0.136 0.673 Group 3 -0.308 0.357 Abbreviations: MPOD = optical density of macular pigment; CDVA = distance visual acuity with correction; L = lutein; Z = zeaxanthin; MZ = mesozeaxanthine; CS = contrast sensitivity; cpd = cycles per degree; GD = glare disability.

Discussion

There was no correlation between increases in visual performance and increases in macular pigment, indicating a neurophysiological effect of macular carotenoids.

Other conclusions: Changes in VP were statistically significant only after 6 months or more

The methods presented this time in contrast sensitivity were at variable spatial frequencies. Low spatial frequencies (for example 1.2 cpd) are indicative of very large objects (for example, a car, a house) while large spatial frequencies (for example, 18 cpd) are indicative of small objects (for example, the letter in a restaurant). The data lead to the following conclusions;

1. The most important effect was on contrast sensitivity, which is one of the most important measures of VP and reflects how the patient actually perceives their own vision.

2. Statistical significance was reached across many spatial frequencies, which means that the detected improvement has implications for general and real-life vision.

3. There was no superior improvement in PV for the Group 2 intervention (ie 10 mg MZ; 10 mg L; 2 mg Z).

Example 4: Effect of two macular carotenoids and a placebo formulation on VP in normal subjects Subjects and recruitment

This study was conducted in 36 normal subjects without AMD. Details of recruitment are given in Example 1. Of the 36 subjects recruited, 32 completed the trial, with one drop-out from each of the intervention groups and two drop-outs from Group 3, the placebo group. All further analysis was confined to those subjects with a complete data set (group 1, n = 11; group 2, n = 11, group 3, n = 10).

Study design and formulations

Normal subjects were divided into 3 groups of 12 subjects (initially) and given the following supplements:

Group 1: L 20; Z 2 mg / day

Group 2: MZ 10; L 10; Z 2 mg / day

Group 3: Placebo 0 mg / day

The carotenoid formulations were in 0.3 ml of vegetable oil and were administered in soft gel capsules.

Visual performance was assessed as described in detail below, at baseline, 3 months, and 6 months.

Statistic analysis

The statistical software package PASW Statistics 18.0 (SPSS Inc., Chicago, Illinois, USA) was used for the analysis. All the quantitative variables investigated showed a typical normal distribution. The means ± SD are presented in the text and tables. Statistical comparisons of the three supplemental groups, at baseline, were performed using one-way ANOVA, while paired-sample t-tests and Repeated measures ANOVA (using a general linear modeling strategy) to analyze visual performance and MPOD measures in each group for supplemental contribution for change across study visits as appropriate. When relevant, the Greenhouse-Geisser correction for sphericity violation was used. A 5% level of significance was used throughout the analysis.

Results

1. Initial analysis

After randomization, one-way analysis of variance revealed no significant differences between groups at baseline, in terms of demographics, macular pigment, visual performance parameters, or other parameters, as illustrated for the selected parameters in the following table 14.

Table 14

V a r ia b le G r u p o 1: M e d ia ± DT G r u p o 2: M e d ia ± D T G r u p o 3: P la c e b o V a lo r P N 12 12 12

Age 56 ± 8 51 ± 13 46 ± 20 0.3 BMI 27 ± 3 25 ± 3 26 ± 5 0.31 BCVA 107 ± 5 109 ± 6 108 ± 6 0.72 MPOD 0.25 0.32 ± 0.13 0.37 ± 0.13 0.35 ± 0.18 0.69 MPOD 0.5 0.25 ± 0.14 0.27 ± 0.12 0.28 ± 0.16 0.88 MPOD 1.0 0 , 15 ± 0.14 0.20 ± 0.07 0.16 ± 0.11 0.46 MPOD 1.75 0.07 ± 0.10 0.10 ± 0.07 0.04 ± 0.04 0, 16 MPOD 3 0.07 ± 0.08 0.08 ± 0.07 0.04 ± 0.05 0.26 SD = standard deviation; BMI = body mass index; BCVA = best visual acuity with correction; MPOD = optical density of macular pigment_________________________________________________________________

2. MPOD response at 3 and 6 months

MPOD measurement

A spatial profile of MPOD was generated at 0.25 °, 0.5 °, 1 °, 1.75 ° and 3 ° of eccentricity of the retina in relation to a reference location of 7 °, using the Macular Densitometer ™, employing a heterochromatic blink photometry (HFP) technique. The subjects were shown an explanatory video of the technique, and a practice session was held before starting the trial. HFP flicker frequencies were optimized following the determination of individual Critical Flicker Fusion (CFF) measurements, in a custom process that optimizes PM measurements (Stringham et al., Exp. Eye res. 2008, 87, 445-453). The MPOD measurement comprised the average of six readings (calculated as the radiance value at which the subject had zero flicker) at each retinal eccentricity, and was considered reliable and acceptable only when the standard deviation of null blink responses was below 0.1.

Table 15: MPOD response and significance to each retinal eccentricity across study visits

Group intervention Initial 3 months Trial 6 months T MRI ANOVA test MPOD 0.25 MPOD 0.25 p * MPOD 0.25 p ** p *** 20 mg L; 2 mg Z 0.32 ± 0.12 0.38 ± 0.15 0.080 0.41 ± 0.14 0.444 0.092 10 mg MZ; 10 mg L ■; 2 mg Z 0.37 ± 0.13 0.49 ± 0.14 0.002 0.50 ± 0.20 0.012 0.002 Placebo 0.35 ± 0.20 0.38 ± 0.20 0.709 0.37 ± 0.18 0.637 0.814 MPOD 0.50 MPOD 0.50 p MPOD 0.50 pp 20 mg L; 2 mg Z 0.27 ± 0.13 0.32 ± 0.22 0.456 0.30 ± 0.14 0.459 0.096 10 mg MZ; 10 mg L ■; 2 mg Z 0.28 ± 0.12 0.38 ± 0.16 0.011 0.37 ± 0.21 0.042 0.010 Placebo 0.28 ± 0.17 0.31 ± 0.16 0.404 0.28 ± 0.16 0.966 0.572 MPOD 1.0 MPOD 1.0 p MPOD 1.0 pp 20 mg L; 2 mg Z 0.16 ± 0.14 0.18 ± 0.12 0.455 0.15 ± 0.14 0.767 0.533 10 mg MZ; 10 mg L ■; 2 mg Z 0.21 ± 0.08 0.28 ± 0.10 0.035 0.27 ± 0.14 0.085 0.047 Placebo 0.16 ± 0.12 0.14 ± 0.11 0.954 0.13 ± 0.10 0.400 0.997 MPOD 1.75 MPOD 1.75 p MPOD 1.75 pp 20 mg L; 2 mg Z 0.08 ± 0.10 0.08 ± 0.10 0.859 0.07 ± 0.10 0.867 0.929 10 mg MZ; 10 mg L ■; 2 mg Z 0.11 ± 0.07 0.19 ± 0.05 0.005 0.18 ± 0.10 0.041 0.036 Placebo 0.03 ± 0.03 0.03 ± 0.05 0.767 0.03 ± 0.05 0.732 0.815 MPOD 3.0 MPOD 3.0 p MPOD 3.0 pp 20 mg L; 2 mg Z 0.05 ± 0.02 0.07 ± 0.06 0.588 0.03 ± 0.03 0.185 0.671 10 mg MZ; 10 mg L ■; 2 mg Z 0.09 ± 0.07 0.11 ± 0.11 0.275 0.10 ± 0.07 0.707 0.915 Placebo 0.02 ± 0.03 0.02 ± 0.03 0.810 0.02 ± 0.05 0.682 0.480 'difference between baseline measurement and 3 months (paired samples T test)

“Difference between baseline measurement and 6 months (paired samples T test)

*** ANOVA of repeated measures across all visits

On this occasion, it can be observed that the greatest increase in MP, at all measured eccentricities, can be observed in group 2, a supplement containing 10 mg of MZ; 10 mg of L; 2 mg of Z.

Visual performance evaluation

Visual acuity (VA) was measured at baseline with a computer generated logMAR test chart (Test Chart 2000 Pro; Thompson Software Solutions, Hatfield, UK) at a viewing distance of 4 m, using the Sloan ETDRS letter set. VA was measured using a one-letter assessment visual acuity rating and was recorded as the average of three measurements provided by the random letter assignment feature of the computer program. The eye with the best visual acuity was chosen as the study eye; however, when both eyes had the same acuity with correction, the right eye was chosen as the study eye.

Contrast sensitivity was measured using a functional acuity contrast test (Optec6500 Vision Tester; Stereo Optical Co. Inc, Chicago, Illinois), incorporating sine wave gratings, presented as Gabor patches, at spatial frequencies of 1.5 , 3, 6, 12 and 18 cycles per degree (cpd) to produce a contrast sensitivity function. The test was carried out under mesopic (3 candela per square meter [cd / m2]) and photopic (85 cd / m2) conditions. (By way of explanation, 3 candela per square meter is considered to represent the upper limit of mesopic conditions: any higher level of illumination is considered to constitute photopic conditions.) The contrast sensitivity test was performed using a Thomson diagram or using the letters EDTRS (Premature Treatment Diabetic Retinopathy Study) in the form of a MAR record at five different spatial frequencies (see, Lorente - Velázquez et al., 2011 Optom. Vis Sci. 88 (10): 1245-1251). Glare disability was assessed using the same test and test conditions, but in the presence of a built-in circumferential LED glare source (42 lux for mesopic testing and 84 lux for photopic glare testing). The LED glare source provided a daylight-simulating color temperature of 6500 ° K, and a spectral emission profile with a single large peak at 453 nm (close to the MP spectral absorbance peak). These assays have been described in more detail elsewhere (Loughman et al. Vis Res.

2010; 50: 1249-1256; Nolan et al. Vis Res. 2011; 51: 459-69). The subject's task and the nature of the test were explained in detail before starting the test, and the subject's performance was closely monitored by an examiner trained during the test, and re-instructed if necessary. Pupil diameter was measured for the background mesopic and photopic conditions used, and also in the presence of both sources of glare using a Neuroptics VIP ™ -200 pupillometer (Neuroptics Inc., Irvine, CA 92612, USA).

The photo-stress recovery time (PRT) of the short wavelength sensitive visual system (SWS) was evaluated using an automated macular photo-stress test (MAP), an adaptation of the Humphrey visual field analyzer (Model 745i Carl Zeiss Meditec Inc Dublin, CA, USA) for the evaluation of the increasing light threshold of the fovea (Dhalla et al., Am J Ophthalmol. 2007; 143 (4), 596-600). To isolate the SWS cones, the medium and long wavelength sensitive cones were desensitized using a three minute sustained exposure to a bleaching background of 100 cd / m2, 570 nm. A Goldmann V stimulus, 440 nm presented for 200 milliseconds was used to test the sensitivity of the SWS system before and after photo-stress. After the three-minute fitting and practice session (during which the subject's performance was assessed for reliability and understanding, the subjects were directed for central fixation between four circumferential light stimuli, and to respond to detection of a "blue" stimulus at that location using the provided response button The sensitivity of the fovea was determined as the average of three consecutive measurements recorded in decibels (dB), with each dB representing a 0.1 logarithmic unit variation in sensitivity. After calculating the initial foveal sensitivity, the subject was exposed to a short wavelength dominated photo-stress stimulus, which consisted of a 5-s exposure to a 300 W lamp viewed at 1 m through a dichroic filter. of low-pass glass, thereby creating a "blue" afterimage of the temporal fovea to mask fixation and reduce fovea sensitivity. After photosostress, a continuous, timed cycle of foveal sensitivity measurements was performed and recorded. The reduction in the sensitivity of the fovea from the beginning was recorded, together with the characteristics of recovery of sensitivity in the SWS system. Again, the pupil diameter was recorded for the background light conditions, and in the presence of the photostress light source.

Ocular light scattering was measured using an Oculus C-Quant (OCULUS Optikgerate GmbH, Wetzlar, Germany), an instrument designed to quantify the effect of light scattering on vision. A central two-part 14 ° test field was visualized in a monocular fashion through the eyepiece of the instrument. Subjects were instructed to respond, using the appropriate response button, to indicate the position of the test half-field on the right or left by blinking more strongly. Subjects were allowed a defined practice session, during which reliable understanding of the task was assessed by the trained examiner. The test results were considered acceptable only when the standard deviation of the measured light scattering value (esd) was <0.08, and the reliability coefficient (Q) was> 1. Absolute light scattering values were recorded. logarithmically [log (s)].

Visual disturbances were assessed during the glare disability and photo-stress test procedures. Subjects were asked to rate their complaints immediately after presentation of the light sources of glare and photostress on a scale ranging from 1-10, where "1" indicated "no ocular discomfort", "5" indicated " moderate eye discomfort "and" 10 "indicated" excruciating eye discomfort ". Such a scale has previously been used effectively in an exemplary macular pigment / glare study (Stringham et al., Invest Ophthalmol Vis Sci. 2011; 52 (10): 7406-15). Visual experience was also assessed by a questionnaire, using a modified version of the visual activities questionnaire, which is used and described in detail elsewhere (Loughman et al. Vis Res. 2010; 50: 1249-1256; Sloane et al. ., The Visual Activities Questionnaire: Developing an instrument for assessing problems in everyday visual tasks. Technical Digest, Noninvasive Assessment of the Visual System, Topical Meeting of the Optical Society of America, January 1992). Iris color was also classified using a standard iris classification scheme as defined by Seddon et al. (Invest Ophthalmol Vis Sci 1990 (31), 8: 1592-1598).

3. BCVA showed no significant effect for any of the intervention groups at 3 months. At 6 months, t-test analysis for paired data revealed a statistically significant improvement in BCVA compared to baseline for group 2 (p = 0.008). The repeated measures ANOVA confirmed a significant change across the three study visits for group 2 (p = 0.034).

4. Contrast sensitivity

Mesopic and photopic contrast sensitivity improved from baseline across a range of spatial frequencies at three months, and particularly at six months. At three months, statistically significant improvements were seen at 1.5 cpd (p = 0.008) for mesopic conditions and at 3 cpd (p = 0.024) and 12 cpd (p = 0.025) for photopic conditions for group 2. At six months, statistically significant improvements in CS were observed across a substantially broader set of spatial frequencies, most notably in mesopic conditions, for group 2, the mesopic CS at 6 cpd significantly improved for group 1 at 6 months (p <0.05). The repeated measures ANOVA confirms the improvements in contrast sensitivity as statistically significant across all study visits for at least 3 of the 5 spatial frequencies tested under mesopic and photopic conditions. A detailed summary of the contrast sensitivity results is provided in Table 16.

Table 16: Change in contrast sensitivity and significance levels at each spatial frequency tested under mesopic and photopic conditions ____________________________________________________ Contrast sensitivity sensitivity group Intervention Contrast ANOVA test at the beginning of six months of Photopic MRT at 1.5 cpd Photopic at 1.5 cpd p * p ** 20 mg L; 2 mg Z 44 ± 26 53 ± 20 0.05 0.12 10 mg MZ; 10 mg L ■; 2 mg Z 49 ± 30 68 ± 28 0.07 0.12 Placebo 52 ± 22 62 ± 29 0.41 0.28 Photopic at 3.0 cpd Photopic at 3.0 cpd

20 mg L; 2 mg Z 85 ± 37 85 ± 29 0.96 0.68 10 mg MZ; 10 mg L ■; 2 mg Z 73 ± 25 100 ± 28 0.002 0.002 Placebo 95 ± 36 94 ± 46 0.84 0.81 Photopic at 6.0 cpd Photopic at 6.0 cpd

20 mg L; 2 mg Z 99 ± 27 100 ± 28 0.71 0.43 10 mg MZ; 10 mg L ■; 2 mg Z 95 ± 36 114 ± 45 0.23 0.26 Placebo 103 ± 54 116 ± 64 0.83 0.88 Photopic at 12.0 cpd Photopic at 12.0 cpd

20 mg L; 2 mg Z 30 ± 10 39 ± 17 0.178 0.26 10 mg MZ; 10 mg L ■; 2 mg Z 32 ± 13 50 ± 30 0.011 0.008 Placebo 57 ± 43 62 ± 42 0.643 0.92 Photopic at 18.0 cpd Photopic at 18.0 cpd

20 mg L; 2 mg Z 8 ± 5 12 ± 9 0.168 0.38 10 mg MZ; 10 mg L ■; 2 mg Z 12 ± 6 23 ± 17 0.059 0.042 Placebo 20 ± 17 17 ± 14 0.527 0.73

Mesopic at 1.5 cpd Mesopic at 1.5 cpd

20 mg L; 2 mg Z 57 ± 30 63 ± 23 0.618 0.83 10 mg MZ; 10 mg L ■; 2 mg Z 52 ± 18 76 ± 24 0.003 0.000 Placebo 65 ± 27 75 ± 24 0.201 0.24 Mesopic at 3.0 cpd Mesopic at 3.0 cpd

20 mg L; 2 mg Z 78 ± 45 74 ± 35 0.792 0.91 10 mg MZ; 10 mg L ■; 2 mg Z 58 ± 17 88 ± 38 0.003 0.001 Placebo 68 ± 39 96 ± 44 0.101 0.11 Mesopic at 6.0 cpd Mesopic at 6.0 cpd

Contrast sensitivity group intervention Contrast ANOVA test at baseline at six months of T MR 20 mg L; 2 mg Z 41 ± 13 53 ± 21 0.06 0.004 10 mg MZ; 10 mg L ■; 2 mg Z 50 ± 19 77 ± 49 0.14 0.058 Placebo 53 ± 46 63 ± 43 0.58 0.82 Mesopic at 12.0 cpd Mesopic at 12.0 cpd

20 mg L; 2 mg Z 7 ± 4 9 ± 6 0.198 0.16 10 mg MZ; 10 mg L ■; 2 mg Z 10 ± 6 33 ± 30 0.040 0.01 Placebo 13 ± 14 21 ± 25 0.400 0.50 Mesopic at 18.0 cpd Mesopic at 18.0 cpd

20 mg L; 2 mg Z 2 ± 0 2 ± 0 NS 0.17 10 mg MZ; 10 mg L ■; 2 mg Z 2 ± 9 11 ± 14 0.047 0.021 Placebo 4 ± 5 5 ± 3 0.593 0.28 RM ANOVA = repeated measures ANOVA across all study visits; NS = not significant (statistical data not calculated as ET of the difference = 0)

* difference between baseline measurement and 6 months (paired samples t-test)

** ANOVA of repeated measures across all visits

Group 1: n = 11; group 2: n = 11; group 3: n = 10

5. Glare disability

Mesopic and photopic glare disability improved from baseline across a range of spatial frequencies at 3 months and 6 months. At three months, statistically significant improvements were observed at 12 cpd (p = 0.048) for mesopic conditions and at 1.5 cpd (p = 0.023) and 3 cpd (p = 0.033) for photopic conditions for group 2. At six months , statistically significant improvements were observed across a substantially broader set of spatial frequencies for group 2. Repeated measures ANOVA across all study visits reveals no statistically significant changes, at any spatial frequency in mesopic glare disability or photopic for groups 1 and 3. Statistically significant improvements in glare disability for group 2, under both mesopic and photopic conditions, for all spatial frequencies tested (other than 18 cpd) were robust for repeated measures ANOVA. A detailed summary of the glare disability results is provided in Table 17.

Table 17: Change in glare disability and significance levels at each spatial frequency tested under mesopic and photopic conditions

Intervention of the group Contrast sensitivity Contrast sensitivity ANOVA test at the beginning at six months of the T MRI Photopic at 1.5 cpd Photopic at 1.5 cpd p * p ** Group 1: 20 mg L; 2 mg Z 56 ± 27 67 ± 20 0.056 0.12 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 50 ± 22 67 ± 22 0.059 0.033 Group 3: Placebo 60 ± 25 74 ± 29 0.134 0.24 Photopic at 3.0 cpd Photopic at 3.0 cpd

Group 1: 20 mg L; 2 mg Z 84 ± 26 95 ± 31 0.175 0.28 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 86 ± 24 121 ± 34 0.003 0.002 Group 3: Placebo 96 ± 30 97 ± 44 0.964 0.92 Photopic at 6.0 cpd Photopic at 6.0 cpd

Group 1: 20 mg L; 2 mg Z 114 ± 43 96 ± 37 0.181 0.26 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 91 ± 39 130 ± 40 0.032 0.04 Group 3: Placebo 105 ± 51 112 ± 58 0.644 0.80 Photopic at 12.0 cpd Photopic at 12.0 cpd

Group 1: 20 mg L; 2 mg Z 34 ± 13 32 ± 14 0.785 0.13 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 42 ± 20 70 ± 25 0.004 0.006 Group 3: Placebo 29 ± 21 62 ± 48 0.06 0.13 Photopic at 18.0 cpd Photopic at 18.0 cpd

Group 1: 20 mg L; 2 mg Z 17 ± 11 23 ± 12 0.35 0.08 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 33 ± 13 65 ± 20 0.17 0.23 Group 3: Placebo 33 ± 15 46 ± 22 0.41 0.75

Mesopic at 1.5 cpd Mesopic at 1.5 cpd

Group 1: 20 mg L; 2 mg Z 23 ± 8 45 ± 35 0.08 0.05 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 39 ± 26 58 ± 29 0.08 0.04 Group 3: Placebo 32 ± 24 38 ± 23 0.76 0.25 Mesopic at 3.0 cpd Mesopic at 3.0 cpd

Group 1: 20 mg L; 2 mg Z 36 ± 10 61 ± 43 0.066 0.06 Group 2: 10 mg MZ; 10 mg L; 2 mg Z 40 ± 14 74 ± 40 0.009 0.02 Group intervention Contrast sensitivity Contrast sensitivity ANOVA test at the beginning at six months of T MRI Group 3: Placebo 54 ± 39 59 ± 46 0.820 0.93 Mesopic at 6.0 cpd Mesopic at 6.0 cpd

Group 1: 20 mg L; 2 mg Z 64 ± 41 90 ± 53 0.15 0.17 Group 2: 10 mg MZ; 10 mg L ■; 2 mg Z 50 ± 19 77 ± 49 0.07 0.049 Group 3: Placebo 53 ± 46 64 ± 43 0.66 0.71 Mesopic at 12.0 cpd Mesopic at 12.0 cpd

Group 1: 20 mg L; 2 mg Z 5 ± 2 10 ± 17 0.303 0.35 Group 2: 10 mg MZ; 10 mg L ■; 2 mg Z 5 ± 2 12 ± 8 0.016 0.014 Group 3: Placebo 7 ± 5 10 ± 7 0.238 0.15 Mesopic at 18.0 cpd Mesopic at 18.0 cpd

Group 1: 20 mg L; 2 mg Z 2 ± 0 2 ± 0 0.34 0.44 Group 2: 10 mg MZ; 10 mg L ■; 2 mg Z 2 ± 1 11 ± 13 0.16 0.21 Group 3: Placebo 4 ± 5 5 ± 3 0.14 0.22 Cpd = cycles per grade

‘Difference between the initial measurement and 6 months (paired samples t-test)

** ANOVA of repeated measures across all visits

Group 1: n = 11; group 2: n = 11; group 3: n = 10

6. Photo-stress recovery time

Photo-stress recovery time did not improve significantly for any of the groups during the study period (p> 0.05 for all). The paired t-test analysis revealed, however, that the improvement in PRT for group 2 (PRT 37 seconds [or 21%] shorter on average at six months compared to baseline) is approximated, but did not reach statistical significance (t = 2.067, p = 0.069).

Measurements of ocular light diffusion did not change significantly for any group (p> 0.05 for all). Visual experience and eye discomfort, determined by questionnaire and discomfort classification, did not change significantly during the study period for any group.

7. Comparison of changes in MP with changes in visual performance parameters

A comparison was made between changes in macular pigment and changes in visual performance parameters between the initial measurement and 6 months. There was no statistically significant relationship between the change in macular pigment and any of the visual performance variables (p> 0.05 for all). Table 18 provides the results for photopic (daytime) and mesopic (night) contrast sensitivity at 1.5 cpd.

Table 18 Changes in macular pigment (at 0.25 ° eccentricity) compared to changes in the following visual performance parameters between baseline and 6 months: BCVA, photopic contrast sensitivity (by day), contrast sensitivity mesopic (at night), photopic contrast sensitivity under glare conditions, mesopic contrast sensitivity under glare conditions.

r p Grupo 1 (20 L, 2 Z) -0,104 0,76 Grupo 2 (10 L, 10 MZ, 2 Z) -0,154 0,651 Grupo 3 (placebo) -0,242 0,5 Cambio en MP frente a cambio en CS mesópica (1,5 cpd) r p Grupo 1 (20 L, 2 Z) 0,394 0,231 Grupo 2 (10 L, 10 MZ, 2 Z) -0,082 0,81 Grupo 3 (placebo) -0,179 0,621 Cambio en MP frente a cambio en GD fotópica (1,5 cpd) r p Grupo 1 (20 L, 2 Z) -0,348 0,294 Grupo 2 (10 L, 10 MZ, 2 Z) 0,263 0,435 Grupo 3 (placebo) 0,331 0,351 Cambio en MP frente a cambio en GD mesópica (1,5 cpd) r p Grupo 1 (20 L, 2 Z) 0,394 0,231 Grupo 2 (10 L, 10 MZ, 2 Z) -0,082 0,81 Grupo 3 (placebo) -0,179 0,621 Abreviaturas: MP = pigmento macular; BCVA = mejor agudeza visual con corrección; L = luteína; Z = zeaxantina; MZ = mesozeaxantina; CS = sensibilidad de contrastes; cpd = ciclos por grado; GD = discapacidad por Cambio en MP frente a cambio en BCVA_______________________________________ r___________ p_____ deslumbramiento.Change in MP versus change in BCVA rp Group 1 (20 L, 2 Z) -0.075 0.827 Group 2 (10 L, 10 MZ, 2 Z) 0.09 0.794 Group 3 (placebo) 0.119 0.743 Change in MP versus change in photopic CS (1.5 cpd)

rp Group 1 (20 L, 2 Z) -0.104 0.76 Group 2 (10 L, 10 MZ, 2 Z) -0.154 0.651 Group 3 (placebo) -0.242 0.5 Change in MP versus change in mesopic CS ( 1.5 cpd) rp Group 1 (20 L, 2 Z) 0.394 0.231 Group 2 (10 L, 10 MZ, 2 Z) -0.082 0.81 Group 3 (placebo) -0.179 0.621 Change in MP versus change in GD photopic (1.5 cpd) rp Group 1 (20 L, 2 Z) -0.348 0.294 Group 2 (10 L, 10 MZ, 2 Z) 0.263 0.435 Group 3 (placebo) 0.331 0.351 Change in MP vs. change in mesopic GD (1.5 cpd) rp Group 1 (20 L, 2 Z) 0.394 0.231 Group 2 (10 L, 10 MZ, 2 Z) -0.082 0.81 Group 3 (placebo) -0.179 0.621 Abbreviations: MP = macular pigment; BCVA = best visual acuity with correction; L = lutein; Z = zeaxanthin; MZ = mesozeaxanthine; CS = contrast sensitivity; cpd = cycles per degree; GD = disability due to Change in MP versus change in BCVA_______________________________________ r___________ p_____ glare.

Surprisingly, these data show that the observed increases in visual performance parameters were independent of increases in macular pigment.

Discussion

In terms of MPOD, there was no significant change in any eccentricity, at 3 or 6 months, in subjects supplemented with a preparation not containing MZ or in subjects administered placebo. In contrast, subjects supplemented with all three macular carotenoids showed a significant increase in MPOD at 4 of the 5 eccentricities tested, at 3 and 6 months.

The present study demonstrates a novel and important effect of increased MP on visual performance among healthy subjects without eye disease. Across a wide range of test modalities and conditions, visual performance improved significantly among subjects showing a significant elevation in MPOD. Specifically, improvements in contrast sensitivity and glare disability (across almost all spatial frequencies, and in daytime and nighttime conditions) and improvements in visual acuity were demonstrated in supplemented subjects with the three macular carotenoids, but no such observations were seen in placebo control subjects or supplemented subjects with L and Z (but not MZ).

The data support the idea that PM can influence visual performance through its optical filtering effects, as the glare disability test protocol includes an LED glare source that displayed a maximum emission profile of length of short wave that matches the known spectral absorbance of MP. The observed improvements in acuity and contrast sensitivity, however, are less consistent with a purely optical explanation. The stimuli used, however, contain a relatively small short-wavelength component. It is possible, therefore, that the increase in MP causes an enhancement of the optical image through a reduction of the detrimental effects of chromatin aberration and light scattering and thereby improves visual acuity. and contrast sensitivity, even for such spectrally broadband stimuli. It is also possible that macular carotenoids, which are intracellular compounds, also play a neurobiological role, thereby contributing to, and / or facilitating, optimal neurophysiological performance and thus visual function (spatial vision limits represent the combined influence of optical and neural efficacy limits). This idea is supported by the observation that there was no correlation between increases in visual performance and increases in macular pigment, suggesting that MP carotenoids may exert effects on visual performance by a neurophysiological mechanism.

In conclusion, a rapid and sustained elevation in MPOD has been demonstrated after supplementation with all three macular carotenoids, and this was not observed in placebo-controlled subjects or in subjects supplemented with a preparation lacking MZ. Furthermore, supplementation with the three macular carotenoids produced significant improvements in contrast sensitivity and glare disability (in photopic or scotopic conditions) and in corrected distance visual acuity, while no such changes were observed in controls with placebo or supplemented subjects with a preparation lacking MZ. These findings have potentially important implications for people involved in activities where optimization of visual significance is important (especially when operating under bright conditions) and warrant further study.

Example 5

Effect of a supplement containing MZ on visual performance in subjects with an atypical distribution of macular pigment (central depression)

Subjects and dosage

Eight subjects with central depressions pre-identified in their macular pigment spatial profile were recruited as described in Example 2 in this study. All eight subjects consumed a supplement containing 10 mg of L, 10 mg of MZ, and 10 mg of Z daily for 3 months.

Methods

Macular pigment optical density (MPOD) was measured as in Example 1 at baseline and after 3 months of supplementation with MZ. Letter contrast sensitivity (Thomsom diagram) was also measured using the method described in Example 3, section 4.

Results

1. MPOD results: As can be seen from table 19 and figure 10, the spatial profile of MP was normalized after supplementation with 10 mg of L, 10 mg of MZ and 10 mg of Z for 3 months. All subjects responded to this intervention. Statistically significant increases were observed at all eccentricities except for 0.5 °.

Table 19.

2. Contrast sensitivity: As can be seen from table 20, there was an improvement in contrast sensitivity after supplementation with 10 mg of L, 10 mg of MZ and 10 mg of Z for 3 months.

Table 20.

Example 6

In one embodiment, the composition of the invention takes the form of a food supplement containing minerals and vitamins, with increased amount of MZ, L and, optionally, Z. The supplement is formulated as a tablet, with the following composition of active ingredients :

MZ 5 mg

L 5 mg

Z 1 mg

Vitamin A 800 micrograms

Thiamine 1.1 mg

Riboflavin 1.4 mg

Vitamin B6 2.0 mg

Vitamin B12 2.5 micrograms

Folic acid 400 micrograms

Niacin 20 mg

Pantothenic acid 6 mg

Biotin 50 micrograms

Vitamin C 80 mg

Vitamin D 20 micrograms

Vitamin E 12 mg

Calcium 120 mg

Magnesium 60 mg

Iron 14 mg

Zinc 10 mg

Copper 1 mg

Iodine 150 micrograms

Manganese 3 mg

Chromium 40 micrograms

Selenium 55 micrograms

Molybdenum 50 micrograms

The following ingredients can be used as a source of the minerals and vitamins.

Minerals: Calcium carbonate, magnesium hydroxide, ferrous fumarate, zinc oxide, copper sulfate, potassium iodide, manganese sulfate, chromium chloride, sodium selenate, sodium molybdate

Vitamins:

Retinyl acetate, thiamine mononitrate, riboflavin, pyridoxine hydrochloride, cyano cobalamin, folic acid, niacin, calcium-D pantothenate, D-biotin, sodium ascorbate, cholecalciferol, D-alpha-tocopherol acetate

Convenient tablets may further comprise one or more of the following fillers: maltodextrin, microcellulose, hydroxypropylmethylcellulose, shellac, talc, acacia, glycerol, titanium dioxide, polyfructose One tablet (eg 500mg) has to be taken per day.

Example 7: Supply of MZ in egg yolks for human consumption

Several workers have shown that L and Z uptake from egg yolks is 2-4 times more effective than from capsules (Handleman eta l., 1999 Am. J. Clin. Nutr. 70, 247-251; Goodrow eta l. ., 2006 J. Nutr. 136, 2519-2524; Johnson 2004, J. Nutr. 134, 1887-1893).

The objective of this study was to feed hens a mixture of L, MZ and Z to determine the total amount of MZ in the yolk. In addition, 24 eggs collected at the end of the experiment by one subject were consumed, one egg / day and the MZ composition of the blood was determined.

Methods

Eight Bovan Goldline hens were obtained at approximately 18 weeks of age

When the hens were producing at least 8 eggs per day in total, the hens were isolated and fed only commercial food. The experiment was started 1 week later when a premix containing the mixed carotenoids was added to the food. The premix provided 250 mg MZ / kg feed with proportions of L 50, MZ 30, Z 20.

Yolk carotenoids were measured in mixes prepared from all eggs collected at baseline, three, and six weeks.

Preparation of egg yolk suspensions

The yolks were weighed individually and mixed with phosphate buffered saline and made up to 50 ml. Two ml of each suspension were mixed in a different universal tube for each of the three separate batches and stored at -40 ° C.

Carotenoid extraction

(i) Egg yolk suspensions

The egg yolk suspension (0.1 ml) was mixed with 0.15 ml of aqueous KOH (25 g / 100 ml of water), 0.15 ml of absolute ethyl alcohol and 0.1 ml of echinenone (internal standard , 0.4 mg / 500 ml of ethyl alcohol) in a glass extraction tube and incubated at 45 ° C for 45 minutes.

The solutions were then chilled and mixed vigorously with 1.5 ml of hexane (containing 500 mg / L BHT) and centrifuged to separate the hexane and aqueous layers. One ml of the upper hexane layer was transferred to an evaporation tube and the residue was re-extracted with 1.5 ml of hexane. After centrifugation, 1.5 ml of the upper layer was removed and the extracts were combined and evaporated to dryness under nitrogen atmosphere at 40 ° C. The residue was made up to 0.15 ml with mobile phase (see Ultracarb HPLC below) and 0.1 ml was injected onto an Ultra Carb column for HPLC analysis.

(ii) Plasma

Blood (10 ml) was collected from a human subject in lithium heparin tubes at baseline, on day 12 and day 24 after consumption of one yolk per day and centrifuged to provide plasma subsequently stored at -40 ° C. . 0.25 ml of plasma was mixed with 0.2 ml of sodium dodecyl sulfate, 0.4 ml of ethyl acetate (internal standard). I know BHT containing hexane (1.0 ml) was added and the mixture was vigorously extracted for 4 min, centrifuged for 10 min and 0.7 ml of the upper hexane layer was removed and evaporated to dryness.

The residue was made up to 0.1 ml with mobile phase (see HPLC procedure below) and 0.05 ml was injected onto the column.

Liquid chromatography (HPLC) to measure L, MZ, Z

Separation and quantification of MZ was achieved using a two column procedure.

Ultracarb procedure: The extracts prepared as described above were reconstituted into a mobile phase comprising acetonitrile: methanol (85:15 containing 0.1% triethylamine). Using the same solvent mixture at 1.5 ml / min, the extracts were subjected to isocratic chromatography using a 3 gm Ultracarb ODS column (250 x 4.6 mm, Phenomenex, UK) and detected using a series detector of photodiodes (model 2996 Waters Ltd) to quantify L and Z MZ at 450 nm. The eluent that coincided with the appearance of MZ Z was collected from the residue line and evaporated to dryness under nitrogen atmosphere.

Chiral chromatography: the Z MZ extract was then reconstituted in 0.1 ml of hexane: isopropanol (90:10) and 50 ul chromatographed on a 10 gm Chiralpak®AD column (250 x 4.6 mm; Chiral Technologies Europe, 67404 Illkirch Cedex, France) to determine the ratio of MZ and Z isomers using gradient elution at 0.8 ml / min starting from 90% hexane and 10% isopropyl alcohol and increasing to 95% hexane in a linear gradient for 30 minutes.

Results

MZ in egg yolks

The mean weights (SD) of the buds at the beginning and end of weeks 3 and 6 were 12.29 (0.35), 14.23 (0.87) and 15.73 (0.72) grams, respectively. . The MZ contents of the buds are shown in table 21. At the beginning only L and Z were present.

The supply of 250 ppm of the carotenoid mixture for 3 weeks produced egg yolks containing 2.78 mg MZ / yolk of which L was around 76%, Z was 13% and MZ was 11%. There was no further increase at 6 weeks.

Plasma

The MZ content in the plasma of a human subject consuming one egg per day is shown in Table 22. The total MZ concentration at baseline was 0.81 gM / l of which L was 53%, Z was a 47% and MZ 0%. The L concentration had almost tripled on day 12, but the concentration then fell to only twice the baseline value on day 24.

The increase in MZ Z on days 12 and 24 was 30% and 23%, respectively, and was solely due to the increase in MZ.

Conclusions

Feeding a mixture of carotenoids to chickens for 3 and 6 weeks increased L MZ Z in egg yolks and in plasma in a subject consuming one egg per day.

The MZ content per yolk rose from about 0.8 mg to 2.8 mg. As the absorption of L and Z from egg yolk is known to be enhanced, two or three eggs from hens fed a mixture of L, Z and MZ could provide enough MZ to improve visual performance in the subject, although this is not known. test.

Table 21. MZ contents of egg yolks from hens fed 250 mg / kg of mixed carotenoids micrograms per yolk _______________________________________________

Weeks L Z MZ Total

0 563 278 0 841

3 2100 366 315 2781

6 2260 328 272 2860

Table 22: MZ contents of plasma in a person consuming one egg per day (units are micromoles per liter),

Day L Z MZ Total

0 0.55 0.26 0 0.81

12 1.20 0.28 0.06 1.54

24 1.06 0.25 0.07 1.38

Example 8: The addition of MZ to dietary and VP formulations

A dry powder formula food supplement composition can be prepared by mixing 5 mg of MZ, 5 mg of L and 1 mg of Z with the contents of 4 sachets containing about 50 g of each of the product of "The Cambridge Diet" , obtained from Cambridge Nutritional Foods Limited, Stafford House, Brakey Road, Corby NN17 5LU, UK (The Cambridge Diet is a registered trademark).

Example 9: Fish oils, MZ and VP

The retina contains a high concentration of Omega 3 fatty acids that are especially abundant in fish oils, for example, oils of salmon, herring, mackerel, anchovies, sardines; also from cril and green-lipped mussels. Omega 3 fatty acids are found as C22.6n-3 eicosapentaenoic acid (EPA) and C22.6n-3 docosahexanoic acid (DHA) and combined make up about 30% of the body oil in fish. The Acceptable Daily Macronutrient Dose (AMND) of EPA DHA is approximately 1.6 g / day for men and 1.1 g / day for women, that is, approximately 5 g and 3.5 g of weighing oil, respectively.

The appearance of a high concentration of Omega 3 fatty acids in the retina suggests that they may play an important role in vision. A combination of macular carotenoids (MC) containing MZ with Omega 3 fatty acids, therefore, would be beneficial for the retina and improve visual performance. The mixture can be in capsules or as an emulsion in a sachet. The latter has the advantage that fewer doses can be given in a sachet while several large capsules (which many elderly people may find difficult to swallow) are needed for NDMA. The emulsion may contain 25-60% fish oils to provide 0.5-2.0 g of Omega 3 fatty acids and enough MC to provide a daily dose of 0.5 mg to 50 mg MC per day .

A commercial preparation of active macular carotenoids (MC) consisting of 10 g mesozeaxanthin, 10 g lutein and 2 g zeaxanthin in 78 ml of cril oil is mixed with 900 ml of salmon oil and manufactured in soft gel capsules containing each a 1 g oil formulation. A daily dose of 5 capsules will provide the 1.5 g of Omega 3 fatty acids and a 22 mg dose of macular carotenoids to improve visual performance.

An alternative embodiment can be formulated as follows:

Ingredients

The mixture of a MC, cril and salmon oil as above 55%

Water 35%

Sucralose (Splenda ™) 4%

Milk powder 5%

Potassium sorbate 0.1%

Alpha tocopherol 0.1%

Flavorings (eg citrus) 0.8%

An emulsion is manufactured in an inert atmosphere using conventional techniques and then packaged in airtight sachets each containing 5 grams of emulsion. The daily dose is 2 sachets per day containing 6 g of Omega 3 fatty acids and 22 mg of MC.

Claims (5)

A non-therapeutic method of improving the visual performance of a human subject without age-related macular degeneration in need of said improvement, the method comprising the step of administering to the subject an effective amount of a composition comprising mesozeaxanthin, lutein and zeaxanthin as a food supplement or food additive, and wherein performing the method improves contrast sensitivity under photopic conditions. A method according to claim 1, wherein the subject consumes the composition at least once a week, preferably at least 3 times a week, and more preferably wherein the subject consumes the composition daily. A method according to claim 1 or 2, wherein the subject consumes the composition, at a desired frequency, for a period of at least 8 weeks, preferably for a period of at least 3 months and more preferably for a period of period of at least 6 months. 4. A method according to any one of claims 1-3, wherein performing the method causes an improvement in the visual performance of a subject having a normal level of macular pigment. A method according to any one of claims 1-3, wherein performing the method causes an improvement in the visual performance of a subject with an atypical "central depression" macular pigment distribution.